Streptavidin alexa fluor 647 conjugate

Manufactured by Thermo Fisher Scientific

Streptavidin Alexa Fluor® 647 Conjugate is a fluorescent-labeled protein that binds to biotin with high affinity. It is used in various bioanalytical techniques that involve biotinylated molecules.

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Lab products found in correlation

Facsaria iiu cytometer, by BD (2 mentions) Imagequant las 4000, by GE Healthcare (1 mentions) Halotag ligand, by Promega (1 mentions) Alexa fluor 660, by Promega (1 mentions) Ivis lumina 3, by PerkinElmer (1 mentions)

Spelling variants of this product

Alexa Fluor 647 (1520 citations) Alexa 647 (424 citations) Streptavidin Alexa Fluor 647 (66 citations) Alexa Fluor conjugates (56 citations) Streptavidin Alexa 647 (28 citations) Alexa Fluors (21 citations) Streptavidin 647 (7 citations)

5 protocols using streptavidin alexa fluor 647 conjugate

ATRX-ADD Binding Assay in HEK293 Cells

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HEK293 cells were lysed in radioimmunoprecipitation assay (RIPA) buffer with protease inhibitors, nicotinamide and trichostatin A. The protein level was quantified by Bradford assay. 10 μg total cell lysates were separated using 12% SDS-PAGE and transferred to polyvinylidene fluoride (PVDF) membrane. After blocking with 5% BSA for 1 hour at room temperature, the membrane was incubated with 100 nM HaloTag ATRX-ADD at 4° for 3 hours. Before incubation, ATRX-ADD was reacted with HaloTag ligand (biotin conjugate or AlexaFluor 660 conjugate) (Promega). For biotin conjugate, the membrane was further incubated with streptavidin-AlexaFluor647 conjugate (Invitrogen) at room temperature for 1 hour. After three washes, the membrane was directly exposed at Cy5 setting (GE ImageQuant LAS 4000).

Su Z., Boersma M.D., Lee J.H., Oliver S.S., Liu S., Garcia B.A, & Denu J.M. (2014). ChIP-less analysis of chromatin states. Epigenetics & Chromatin, 7, 7.

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Insulin Binding Titration Assay

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The 3×FLAG-mIR cell line and HEK-Cre+ cells were assessed via an IR binding titration assay. Cells were incubated with various sample concentrations of biotinylated-insulin (Dr. Finn, University of Pittsburg, PA) ranging from 1 × 10^–7 M to 1 × 10^–13 M, for 45 min at 4°C. Cells were stained with Streptavidin Alexa Fluor^® 647 Conjugate (Invitrogen) secondary reagent and analyzed on a BD FACSAriaIIu cytometer.

Morran M.P., Al-Dieri A.G., Nestor-Kalinoski A.L., Jordan R.K., Gupta N.K, & McInerney M.F. (2018). Insulin receptor based lymphocyte trafficking in the progression of type 1 diabetes. Journal of Biological Methods, 5(1), e85.

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Insulin Binding Affinity Titration Assay

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The 3×FLAG-mIR cell line and HEK-Cre+ cells were assessed via an IR binding titration assay. Cells were incubated with various sample concentrations of biotinylated-insulin (Dr. Finn, University of Pittsburg, PA) ranging from 1 × 10⁻⁷ M to 1 × 10⁻¹³ M, for 45 min at 4°C. Cells were stained with Streptavidin Alexa Fluor^® 647 Conjugate (Invitrogen) secondary reagent and analyzed on a BD FACSAriaIIu cytometer.

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Quantifying Shh-PTCH1 Interactions

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293 cells were transiently transfected with GFP-tagged Ptch1 constructs. After 24 hours, cells were dissociated using 10 mM EDTA, washed with HPBS 0.5 mM Ca²⁺, and pelleted by centrifugation. Cells were then resuspended in binding buffer (HPBS, 0.5 mM Ca²⁺, 0.5 mg/ml BSA) and incubated with purified ShhN-biotin (1:400 dilution) for 30 minutes at 4° C. Cells were then washed three times in binding buffer by centrifugation and subsequently incubated with Alexa Fluor 647 streptavidin conjugate (Invitrogen) for 15 minutes at 4° C. Cells were then washed three times by centrifugation in wash buffer (binding buffer plus 1 mM biotin) and the percentages of cells bound by ShhN were quantified by flow cytometry after gating for PTCH1-GFP expression (BD FACSAria II, Stanford Stem Cell Institute FACS Core).

Zhang Y., Bulkley D.P., Xin Y., Roberts K.J., Asarnow D.E., Sharma A., Myers B.R., Cho W., Cheng Y, & Beachy P.A. (2018). Structural basis for cholesterol transport-like activity of the Hedgehog receptor Patched. Cell, 175(5), 1352-1364.e14.

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In Vivo Fluorescence Imaging of Antigen Localization

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To assess the localization of antigens after their application, the anesthetized mice were injected with 5 μg of Alexa Fluor® 647-streptavidin conjugate (Invitrogen) via i.n. or s.l. route. The distribution of the fluorescence dye was monitored by in vivo imaging system (IVIS Lumina III; PerkinElmer Health Sciences, Waltham, MA) with Ex/Em of 640 nm/710 nm at 0.5, 1, and 24 h following injection. The average photon radiance on the certain surface of a mice was expressed as photons per second per centimeter squared per steradian (p/s/cm²/sr).

Cheon I.S., Kim J.Y., Choi Y., Shim B.S., Choi J.A., Jung D.I., Kim J.O., Braciale T.J., Youn H., Song M.K, & Chang J. (2019). Sublingual Immunization With an RSV G Glycoprotein Fragment Primes IL-17-Mediated Immunopathology Upon Respiratory Syncytial Virus Infection. Frontiers in Immunology, 10, 567.

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